Our objective is to determine scientifically if Therapeutic Touch is able to affect bone formation in human osteoblast cultures in vitro. Therapeutic Touch is a non-invasive healing modality and it has been used for diverse medical problems. However, there is doubt about its biological plausibility, therefore, rigorous methods are needed to test its value. Therapeutic Touch has been used mostly on patients. However, we propose to investigate its effect on bone growth in vitro, which is a more simple system, amenable to more exacting scientific evaluation. Cell adhesion, intracellular signaling, adhesion, morphology, proliferation and differentiation will be assessed. We will employ the same Therapeutic Touch practitioner to affect changes in bone healing in patients in vivo (Project 1) as bone growth in vitro. Our long range goals are to understand the biologic basis for effects on bone growth by Therapeutic Touch, and then utilize these findings to improve bone healing in vivo. Our specific aims are to investigate the effect of Therapeutic Touch on the growth, differentiation and mineralization of human osteoblast culture. Cell proliferation, collagen synthesis and calcification will be analyzed in human osteoblast cultures from middle-aged patients. We will also examine any differences in the cytoskeleton and integrin levels in the human osteoblast cultures, which have been exposed to Therapeutic Touch. Integrin levels will be assayed by Western and Northern blot analysis. Immunofluorescence microscopy of the cytoskeleton, integrins and cell morphology will also be performed. If differences are found in integrin levels and/or the cytoskeleton then the MAP kinase signaling pathway leading to the activation of AP-1 transcription factors will be assayed. Western blot analysis of specific signaling factors, their phosphorylation levels and the levels of AP-1 transcription factors, which are responsible for osteoblast growth and differentiation, will be examined. Our goal is to develop novel strategies to stimulate bone formation in fracture repair and/or musculoskeletal diseases, and to develop a system(s) in which the biologic basis for Therapeutic Touch can be studied.